Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1993 Nov 1;178(5):1825–1830. doi: 10.1084/jem.178.5.1825

Thromboxane A2 receptor is highly expressed in mouse immature thymocytes and mediates DNA fragmentation and apoptosis

PMCID: PMC2191260  PMID: 8228829

Abstract

We have recently revealed that the thymus is the organ showing the highest expression of thromboxane (TX) A2 receptor in mice. In this study, thymic cell populations expressing the receptor were identified, and the effects of a TXA2 agonist on these cells were examined. Radioligand binding using a TXA2 receptor-specific radioligand revealed a single class of binding sites in the thymocytes with an affinity and specificity identical to those reported for the TXA2 receptor. The receptor density in these cells was comparable to that seen in blood platelets. This receptor is most highly expressed in CD4-8- and CD4+8+ immature thymocytes, followed by CD4+8- and CD4-8+ cells. The receptor density in splenic T cells was less than one fifth of that in CD4+8+ cells and no binding activity was detectable in splenic B cells. The addition of a TXA2 agonist, STA2, to thymocytes induced the disappearance of the CD4+8+ cells in a time- and concentration- dependent manner and caused DNA fragmentation. These changes were blocked by a specific TXA2 antagonist, S-145. These results demonstrate that TXA2 induces apoptotic cell death in immature thymocytes by acting on the TXA2 receptor on their cell surface and suggest a role for the TXA2/TXA2 receptor system in the thymic micro-environment.

Full Text

The Full Text of this article is available as a PDF (596.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ackerman R. C., Murdoch W. J. Prostaglandin-induced apoptosis of ovarian surface epithelial cells. Prostaglandins. 1993 May;45(5):475–485. doi: 10.1016/0090-6980(93)90123-o. [DOI] [PubMed] [Google Scholar]
  2. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brass L. F., Shaller C. C., Belmonte E. J. Inositol 1,4,5-triphosphate-induced granule secretion in platelets. Evidence that the activation of phospholipase C mediated by platelet thromboxane receptors involves a guanine nucleotide binding protein-dependent mechanism distinct from that of thrombin. J Clin Invest. 1987 Apr;79(4):1269–1275. doi: 10.1172/JCI112947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Giunta M., Favre A., Ramarli D., Grossi C. E., Corte G. A novel integrin involved in thymocyte-thymic epithelial cell interactions. J Exp Med. 1991 Jun 1;173(6):1537–1548. doi: 10.1084/jem.173.6.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Goldyne M. E. Lymphocytes and arachidonic acid metabolism. Prog Allergy. 1988;44:140–152. [PubMed] [Google Scholar]
  6. Halushka P. V., Mais D. E., Garvin M. Binding of a thromboxane A2/prostaglandin H2 receptor antagonist to guinea-pig platelets. Eur J Pharmacol. 1986 Nov 12;131(1):49–54. doi: 10.1016/0014-2999(86)90514-5. [DOI] [PubMed] [Google Scholar]
  7. Hanasaki K., Nakano K., Kasai H., Kurihara H., Arita H. Identification of thromboxane A2 receptor in cultured vascular endothelial cells of rat aorta. Biochem Biophys Res Commun. 1988 Mar 30;151(3):1352–1357. doi: 10.1016/s0006-291x(88)80511-4. [DOI] [PubMed] [Google Scholar]
  8. Hanasaki K., Nakano T., Arita H. Receptor-mediated mitogenic effect of thromboxane A2 in vascular smooth muscle cells. Biochem Pharmacol. 1990 Dec 1;40(11):2535–2542. doi: 10.1016/0006-2952(90)90096-4. [DOI] [PubMed] [Google Scholar]
  9. Hirata M., Hayashi Y., Ushikubi F., Yokota Y., Kageyama R., Nakanishi S., Narumiya S. Cloning and expression of cDNA for a human thromboxane A2 receptor. Nature. 1991 Feb 14;349(6310):617–620. doi: 10.1038/349617a0. [DOI] [PubMed] [Google Scholar]
  10. Hockenbery D. M., Zutter M., Hickey W., Nahm M., Korsmeyer S. J. BCL2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6961–6965. doi: 10.1073/pnas.88.16.6961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hockenbery D., Nuñez G., Milliman C., Schreiber R. D., Korsmeyer S. J. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature. 1990 Nov 22;348(6299):334–336. doi: 10.1038/348334a0. [DOI] [PubMed] [Google Scholar]
  12. Homo-Delarche F., Duval D., Papiernik M. Prostaglandin production by phagocytic cells of the mouse thymic reticulum in culture and its modulation by indomethacin and corticosteroids. J Immunol. 1985 Jul;135(1):506–512. [PubMed] [Google Scholar]
  13. Honda A., Sugimoto Y., Namba T., Watabe A., Irie A., Negishi M., Narumiya S., Ichikawa A. Cloning and expression of a cDNA for mouse prostaglandin E receptor EP2 subtype. J Biol Chem. 1993 Apr 15;268(11):7759–7762. [PubMed] [Google Scholar]
  14. Itoh N., Yonehara S., Ishii A., Yonehara M., Mizushima S., Sameshima M., Hase A., Seto Y., Nagata S. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell. 1991 Jul 26;66(2):233–243. doi: 10.1016/0092-8674(91)90614-5. [DOI] [PubMed] [Google Scholar]
  15. Katsura M., Miyamoto T., Hamanaka N., Kondo K., Terada T., Ohgaki Y., Kawasaki A., Tsuboshima M. In vitro and in vivo effects of new powerful thromboxane antagonists (3-alkylamino pinane derivatives). Adv Prostaglandin Thromboxane Leukot Res. 1983;11:351–357. [PubMed] [Google Scholar]
  16. Kujubu D. A., Fletcher B. S., Varnum B. C., Lim R. W., Herschman H. R. TIS10, a phorbol ester tumor promoter-inducible mRNA from Swiss 3T3 cells, encodes a novel prostaglandin synthase/cyclooxygenase homologue. J Biol Chem. 1991 Jul 15;266(20):12866–12872. [PubMed] [Google Scholar]
  17. Mazda O., Watanabe Y., Gyotoku J., Katsura Y. Requirement of dendritic cells and B cells in the clonal deletion of Mls-reactive T cells in the thymus. J Exp Med. 1991 Mar 1;173(3):539–547. doi: 10.1084/jem.173.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McConkey D. J., Hartzell P., Nicotera P., Orrenius S. Calcium-activated DNA fragmentation kills immature thymocytes. FASEB J. 1989 May;3(7):1843–1849. doi: 10.1096/fasebj.3.7.2497041. [DOI] [PubMed] [Google Scholar]
  19. McConkey D. J., Orrenius S., Jondal M. Cellular signalling in programmed cell death (apoptosis). Immunol Today. 1990 Apr;11(4):120–121. doi: 10.1016/0167-5699(90)90048-e. [DOI] [PubMed] [Google Scholar]
  20. McCormack J. E., Kappler J., Marrack P., Westcott J. Y. Production of prostaglandin E2 and prostacyclin by thymic nurse cells in culture. J Immunol. 1991 Jan 1;146(1):239–243. [PubMed] [Google Scholar]
  21. Namba T., Sugimoto Y., Hirata M., Hayashi Y., Honda A., Watabe A., Negishi M., Ichikawa A., Narumiya S. Mouse thromboxane A2 receptor: cDNA cloning, expression and northern blot analysis. Biochem Biophys Res Commun. 1992 May 15;184(3):1197–1203. doi: 10.1016/s0006-291x(05)80009-9. [DOI] [PubMed] [Google Scholar]
  22. Narumiya S., Okuma M., Ushikubi F. Binding of a radioiodinated 13-azapinane thromboxane antagonist to platelets: correlation with antiaggregatory activity in different species. Br J Pharmacol. 1986 Jun;88(2):323–331. doi: 10.1111/j.1476-5381.1986.tb10208.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nüsing R., Sauter G., Fehr P., Dürmüller U., Kasper M., Gudat F., Ullrich V. Localization of thromboxane synthase in human tissues by monoclonal antibody Tü 300. Virchows Arch A Pathol Anat Histopathol. 1992;421(3):249–254. doi: 10.1007/BF01611182. [DOI] [PubMed] [Google Scholar]
  24. Nüsing R., Ullrich V. Immunoquantitation of thromboxane synthase in human tissues. Eicosanoids. 1990;3(3):175–180. [PubMed] [Google Scholar]
  25. Ogletree M. L. Overview of physiological and pathophysiological effects of thromboxane A2. Fed Proc. 1987 Jan;46(1):133–138. [PubMed] [Google Scholar]
  26. Pollock W. K., Armstrong R. A., Brydon L. J., Jones R. L., MacIntyre D. E. Thromboxane-induced phosphatidate formation in human platelets. Relationship to receptor occupancy and to changes in cytosolic free calcium. Biochem J. 1984 May 1;219(3):833–842. doi: 10.1042/bj2190833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ruiz P., Rey L., Spurney R., Coffman T., Viciana A. Thromboxane augmentation of alloreactive T cell function. Transplantation. 1992 Sep;54(3):498–505. doi: 10.1097/00007890-199209000-00021. [DOI] [PubMed] [Google Scholar]
  28. Smith C. A., Williams G. T., Kingston R., Jenkinson E. J., Owen J. J. Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature. 1989 Jan 12;337(6203):181–184. doi: 10.1038/337181a0. [DOI] [PubMed] [Google Scholar]
  29. Sugimoto Y., Namba T., Honda A., Hayashi Y., Negishi M., Ichikawa A., Narumiya S. Cloning and expression of a cDNA for mouse prostaglandin E receptor EP3 subtype. J Biol Chem. 1992 Apr 5;267(10):6463–6466. [PubMed] [Google Scholar]
  30. Swat W., Ignatowicz L., von Boehmer H., Kisielow P. Clonal deletion of immature CD4+8+ thymocytes in suspension culture by extrathymic antigen-presenting cells. Nature. 1991 May 9;351(6322):150–153. doi: 10.1038/351150a0. [DOI] [PubMed] [Google Scholar]
  31. Tippetts M. T., Varnum B. C., Lim R. W., Herschman H. R. Tumor promoter-inducible genes are differentially expressed in the developing mouse. Mol Cell Biol. 1988 Oct;8(10):4570–4572. doi: 10.1128/mcb.8.10.4570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ushikubi F., Nakajima M., Hirata M., Okuma M., Fujiwara M., Narumiya S. Purification of the thromboxane A2/prostaglandin H2 receptor from human blood platelets. J Biol Chem. 1989 Oct 5;264(28):16496–16501. [PubMed] [Google Scholar]
  33. Watanabe Y., Mazda O., Aiba Y., Iwai K., Gyotoku J., Ideyama S., Miyazaki J., Katsura Y. A murine thymic stromal cell line which may support the differentiation of CD4-8- thymocytes into CD4+8- alpha beta T cell receptor positive T cells. Cell Immunol. 1992 Jul;142(2):385–397. doi: 10.1016/0008-8749(92)90299-5. [DOI] [PubMed] [Google Scholar]
  34. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]
  35. Yonehara S., Ishii A., Yonehara M. A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor. J Exp Med. 1989 May 1;169(5):1747–1756. doi: 10.1084/jem.169.5.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES